【摘要】 目的　观察分泌型aFGF真核表达载体转染大鼠心肌细胞后 ,aFGF在体内、外的分泌表达及转染心肌细胞的移植对宿主心肌血管形成作用的影响。方法　构建分泌型aFGF真核表达载体(pCDNA3 1+SP aFGF) ,转染培养大鼠的心肌细胞 ,检测体外心肌细胞对aFGF的分泌表达 ;将转染细胞移植入同种大鼠心肌 ,检测移植细胞在体内对aFGF的表达及宿主心肌的血管生成。结果　硝酸纤维素膜点杂交和ELISA显示 ,aFGF在转染 2 4～ 96h细胞培养上清中有较高的表达 ;接受转染心肌细胞移植96h后免疫组化检查 ,发现aFGF在受体心肌内得到分泌表达 ,2 0d后组织切片发现 ,移植心肌细胞周围的心肌中有较多的新生毛细血管。结论　转染分泌型aFGF真核表达载体的心肌细胞 ,在体内、外能分泌表达aFGF ;通过心肌细胞移植 ,分泌的aFGF在受体心肌组织中有较强的血管生成作用更多还原

【Abstract】 Objective: aFGF is a potent angiogenic polypeptide, the structure of which lacks a signal peptide sequence for secretion . The purpose of present study is to study the expression of constructed secretory aFGF gene in cardiomyocytes and its angiogenic effect on host myocardium. Methods: Secretory eucaryotic expression vector pCDNA3.1+SPaFGF was constructed and transfected in vitro to the neonate rat cardiomyocytes. The expression of aFGF was assessed .After transplantation of cells transfected with aFGF into the hearts of adult syngenic mice,the expression of aFGF in vivo and its angiogenic effect in host myocardium were studied.Results: The expression of aFGF was demonstrated by Western blotting and ELISA in the cultured media 24-96 hours later. Using immunohistochemistry analysis, we detected aFGF in the recipient myocardium 96 hours later. 20 days after cell transplantation, histological study revealed more blood vessels around the grafted cardiomyocytes in the host myocardium which was not marked in the controlled myocardium. Conclusions: The plasmid we constructed succeeds in expressing and secreting aFGF peptide possibly leading to angiogenesis in the recipient myocardium.更多还原